目的 基于高效液相色谱-四极杆飞行时间串联质谱法（UPLC-Q-TOF-MS/MS）分析五味子Schisandra chinensis藤茎的主要成分，并结合网络药理学和分子对接方法预测保肝药效物质和潜在的作用靶点及通路。方法 根据UPLC-Q-TOF-MS/MS二级质谱裂解碎片信息，并结合文献数据，对五味子藤茎化学成分进行分析鉴定；通过中药系统药理学数据库和分析平台（Traditional Chinese Medicine Systems Pharmacology Database and Analysis Platform，TCMSP），从鉴定获得的化学成分中筛选出口服生物利用度（oral bioavailability，OB）≥30%且符合类药五原则的活性成分；运用TCSMP数据库和SwissTargetPrediction数据库查找和预测五味子藤茎的成分靶点；通过在线人类孟德尔遗传数据库（Online Mendelian Inheritance in Man，OMIM）、GeneCards数据库和京都基因与基因组百科全书（Kyoto encyclopedia of genes and genomes，KEGG）数据库筛选疾病靶点。采用String11.5数据库和Cytoscape 3.7.2软件构建PPI网络，并筛选核心靶点。利用DAVID 6.8进行基因本体（gene ontology，GO）注释和KEGG通路富集分析，并通过AutoDockTools 1.5.6软件对五味子藤茎的活性成分与作用靶点进行分子对接验证。构建酒精致肝细胞损伤体外模型，并考察五味子藤茎提取物对肝损伤细胞的保护作用。结果 从五味子藤茎中共鉴定出40个主要成分，包括木脂素类、黄酮类、有机酸类化合物；进一步筛选出活性成分12个，活性成分与疾病交集靶点118个；通过网络拓扑筛选获得核心靶点16个；富集分析发现五味子藤茎保肝作用主要涉及的通路包括：肿瘤坏死因子（tumor necrosis factor，TNF）、丙型肝炎（hepatitis C）、乙型肝炎（hepatitis B）、血管内皮生长因子（vascular endothelial growth factor，VEGF）信号通路等。分子对接结果表明，五味子甲素、五味子乙素、五味子丙素、五味子酯甲、槲皮素、戈米辛M2、戈米辛R等成分可与表皮生长因子（epidermal growth factor receptor，EGFR）、磷脂酰肌醇-3-激酶催化亚基α（phosphatidylinositol-3-kinase catalytic subunit alpha isoform，PIK3CA）、TNF、人半胱氨酸-天冬氨酸蛋白酶3（cysteine aspartic protease-3，Caspase-3）等靶点蛋白发生相互作用，结合活性良好。五味子藤茎可抑制酒精引起的人正常肝细胞系L-02存活率降低，降低谷丙转氨酶（alanine aminotransferase，ALT）和谷草转氨酶（aspartate aminotransferase，AST）活性。结论 五味子藤茎具有保肝作用，主要药效物质为木脂素类，可通过多靶点、多通路发挥保肝作用。

Objective To analyze the main components of Schisandra chinensis rattan stems (SCRS) by ultra performance liquid chromatography-quadrupole time-of-flight mass spectrometry (UPLC-Q-TOF-MS/MS), and predict the hepatoprotective substances and potential targets and signaling pathways by network pharmacology and molecular docking. Methods According to the fragmentation of secondary mass spectrometry and literature data, the chemical constituents of SCRS were analyzed and identified. The active components with OB ≥ 30% in accordance with the five principles of drugs were screened through the TCMSP database. The potential targets of active components of SCRS were checked in TCMSP database and predicted by SwissTargetPrediction online platform. The “hepatitis” and “liver injury” were selected as key words to screen disease targets with OMIM, GeneCards and KEGG databases. PPI network was constructed using String11.5 database and Cytoscape 3.7.2 software, and core targets were screened. DAVID 6.8 database was used for gene ontology (GO) anotation and KEGG pathway enrichment analysis. AutoDockTools 1.5.6 software was used to verify the molecular docking between the active components and the key targets of SCRS. The in vitro model of liver cell injury was established and the protective effect of SCRS extract on liver cell injury was investigated. Results A total of 40 main components were identified in SCRS, including lignans, flavonoids, organic acids and so on. 12 active ingredients were further screened, and 118 intersection targets of active ingredients and diseases were obtained. Sixteen core targets were obtained by network topology screening. Enrichment analysis showed that the main pathways involved in the hepatoprotective effect of SCRS included TNF, hepatitis C, hepatitis B and VEGF signaling pathways. The results of molecular docking showed that ingredients such as schisandrin A, schisandrin B, schisandrin C, schisantherin A, quercetin, gomixin M2 and gomixin R could interact with targets including EGFR, PIK3CA, TNF, Caspase-3 and other proteins with good binding affinity. SCRS could suppress the decrease of L-02 cell survival rate and inhibit ALT and AST activity induced by alcohol. Conclusion SCRS shows a good hepatoprotective effect, and the main medicinal substance is lignans, which can exert hepatoprotective effect through multi-target and multi-pathway.

R284.1；R285

黑龙江省重点研发计划指导类项目（GZ20210110）；黑龙江省博士后科研启动金资助项目（LBH-Q16133）；哈尔滨商业大学青年创新人才支持计划项目（2020CX11）；哈尔滨市科技攻关计划项目（2012DB3BS057）